The Division of the National Toxicology Program (DNTP) develops and carries out programs in medium and high throughput screening (HTS) of environmental substances utilizing new approach methodologies for rapid detection of biological activities of significance to toxicology. The Biomolecular Screening Branch (BSB) specifically administers the NTP HTS program initiative to implement its vision for toxicology in the 21st century; this vision places an increased emphasis on the use of these new alternative methods for targeting key pathways, molecular events, or processes linked to disease or injury, in an attempt to link exposure-induced activities with human physiologically-relevant effects. The both the NTP Laboratory (NTPLab) and the Toxicology Branch evaluate and implement new approach methodologies for critical areas of NTP concern, developing assays and approaches where needed. The NTP Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM) supports the development and evaluation of new, revised and alternative methods to identify hazards to human health and the environment, with a focus on replacing, reducing, or refining animal use. The ultimate goal of the NTP effort in utilizing new approach methodologies is to better evaluate the human health impact of environmental exposures quickly and efficiently by incorporating these new alternative approaches into a research and testing framework for translational toxicology in the 21st century.